Gibellini Davide, Re Maria Carla, La Placa Michele, Zauli Giorgio
Department of Clinical and Experimental Medicine, St Orsola Hospital, University of Bologna, Via Massarenti 9, 40138, Bologna, Italy.
Virus Res. 2002 Dec;90(1-2):337-45. doi: 10.1016/s0168-1702(02)00253-8.
Several studies have indicated that human immunodeficiency virus type-1 (HIV-1) transactivating Tat protein is essential for proviral DNA transcription and virus replication. In addition, it is actively released from acutely HIV-1-infected cells and interacts either with the same virus-infected and virus producing cell, or with bystander uninfected cells, influencing the expression of several genes and related cellular functions. The main goal of this paper was to determine the Tat-related expression of basic cellular genes in a permanently tat transfected CD4+ cell line, to identify the cellular genes influenced by the presence of endogenous-exogenous Tat protein. For this purpose, we analyzed, by a cDNA-membrane-array assay, cellular mRNAs expressed in serum-free cultures of lymphoblastoid CD4(+) Jurkat cells, stably transfected with a plasmid constitutively expressing tat gene, in comparison with Jurkat cells transfected with the backbone plasmid only, and parental Jurkat cells. The expression of mRNAs in permanently tat-transfected Jurkat cells showed significant differences in 24 out of 1176 analyzed genes in comparison with parental or backbone plasmid transfected cells. Most of the genes overexpressed in permanently tat-transfected Jurkat cells, belong to transcription factors, or to receptors, adaptors, and mediators of signal transduction pathways, and to factors involved in response to oxidative stress, suggesting a complex regulation of CD4(+) T-lymphoid cell survival and proliferation by HIV-1 Tat protein.
多项研究表明,人类免疫缺陷病毒1型(HIV-1)反式激活Tat蛋白对于前病毒DNA转录和病毒复制至关重要。此外,它可从急性HIV-1感染的细胞中主动释放出来,并与相同的病毒感染及病毒产生细胞相互作用,或与旁观者未感染细胞相互作用,影响多个基因的表达及相关细胞功能。本文的主要目的是确定在稳定转染tat基因的永生化CD4+细胞系中与Tat相关的基本细胞基因表达情况,以识别受内源性-外源性Tat蛋白存在影响的细胞基因。为此,我们通过cDNA膜阵列分析,比较了稳定转染组成型表达tat基因质粒的淋巴母细胞样CD4(+) Jurkat细胞在无血清培养中表达的细胞mRNA,以及仅转染空载体质粒的Jurkat细胞和原始Jurkat细胞。与原始或转染空载体质粒的细胞相比,在稳定转染tat基因的Jurkat细胞中,1176个分析基因中有24个基因的mRNA表达存在显著差异。在稳定转染tat基因的Jurkat细胞中过度表达的大多数基因,属于转录因子、信号转导途径的受体、衔接蛋白和介质,以及参与氧化应激反应的因子,这表明HIV-1 Tat蛋白对CD4(+) T淋巴细胞的存活和增殖具有复杂的调控作用。